Wang Tingting, Nielsen Kirstine L, Frisch Kim, Lassen Johan K, Nielsen Camilla B, Andersen Charlotte U, Villesen Palle, Andreasen Mette F, Hasselstrøm Jørgen B, Johannsen Mogens
Department of Forensic Medicine, Section for Forensic Chemistry, Aarhus University, Aarhus, Denmark.
Bioinformatics Research Centre, Aarhus University, Aarhus, Denmark.
Front Pharmacol. 2022 Mar 3;13:816376. doi: 10.3389/fphar.2022.816376. eCollection 2022.
GHB is an endogenous short-chain organic acid presumably also widely applied as a rape and knock out drug in cases of drug-facilitated crimes or sexual assaults (DFSA). Due to the endogenous nature of GHB and its fast metabolism , the detection window of exogenous GHB is however narrow, making it challenging to prove use of GHB in DFSA cases. Alternative markers of GHB intake have recently appeared though none has hitherto been validated for forensic use. UHPLC-HRMS based screening of blood samples for drugs of abuse is routinely performed in several forensic laboratories which leaves an enormous amount of unexploited data. Recently we devised a novel metabolomics approach to use archived data from such routine screenings for elucidating both direct metabolites from exogenous compounds, but potentially also regulation of endogenous metabolism and metabolites. In this paper we used UHPLC-HRMS data acquired over a 6-year period from whole blood analysis of 51 drivers driving under the influence of GHB as well as a matched control group. The data were analyzed using a metabolomics approach applying a range of advanced analytical methods such as OPLS-DA, LASSO, random forest, and Pearson correlation to examine the data in depth and demonstrate the feasibility and potential power of the approach. This was done by initially detecting a range of potential biomarkers of GHB consumption, some that previously have been found in controlled GHB studies, as well as several new potential markers not hitherto known. Furthermore, we investigate the impact of GHB intake on human metabolism. In aggregate, we demonstrate the feasibility to extract meaningful information from archived data here exemplified using GHB cases. Hereby we hope to pave the way for more general use of the principle to elucidate human metabolites of e.g. new legal or illegal drugs as well as for applications in more global and large scale metabolomics studies in the future.
γ-羟基丁酸（GHB）是一种内源性短链有机酸，据推测在药物辅助犯罪或性侵犯（DFSA）案件中也被广泛用作强奸和迷昏药物。然而，由于GHB的内源性及其快速代谢，外源性GHB的检测窗口期很窄，这使得在DFSA案件中证明使用了GHB具有挑战性。尽管最近出现了GHB摄入的替代标志物，但迄今为止尚无一种经过法医学验证。基于超高效液相色谱-高分辨率质谱（UHPLC-HRMS）的血液样本滥用药物筛查在多个法医学实验室中常规进行，这留下了大量未利用的数据。最近，我们设计了一种新的代谢组学方法，利用此类常规筛查的存档数据来阐明外源性化合物的直接代谢物，以及潜在的内源性代谢和代谢物的调节。在本文中，我们使用了6年期间从51名受GHB影响驾驶的司机以及一个匹配对照组的全血分析中获取的UHPLC-HRMS数据。使用代谢组学方法对数据进行分析，应用了一系列先进的分析方法，如正交偏最小二乘法判别分析（OPLS-DA）、套索回归（LASSO）、随机森林和皮尔逊相关性，以深入检查数据并证明该方法的可行性和潜在能力。这是通过首先检测一系列GHB消费的潜在生物标志物来完成的，其中一些先前已在受控GHB研究中发现，以及一些迄今未知的新潜在标志物。此外，我们研究了GHB摄入对人体代谢的影响。总体而言，我们证明了从存档数据中提取有意义信息的可行性，这里以GHB案例为例进行了说明。借此，我们希望为更广泛地使用该原理来阐明例如新的合法或非法药物的人体代谢物以及未来在更全面和大规模的代谢组学研究中的应用铺平道路。
